Fig 1: Glp2r inhibition negates aberrant gut glucose sensing and exacerbates gut glucose absorption upon high fructose intake. (A) Mice consuming diets rich in carbohydrates either from glucose or with a portion of glucose replaced by fructose received daily injections with the glucagon-like peptide (Glp)2 receptor (Glp2r) antagonist Glp2 (3–33) for 23 consecutive days. (B) At the beginning of week 7, mice were gavaged with the non-metabolizable glucose analog 3-O-methylglucose (3-OMG, 4 mg/mouse) and the area under 3-OMG excursion curves was calculated to assess changes in gut glucose absorption. (C, D) At the conclusion of the 7-week study protocol (day 52), proximal and distal small intestine sections were collected for gene expression analysis of sugar transporters. (E) Gut morphometry was performed in a blinded fashion by two operators on H&E-stained mid-third jejunum sections. Representative images of proximal and distal areas of the mid-jejunum and quantification of villus height (v.h.), villus width (v.w.), and crypt depth (c.d.) in these gut segments were analyzed. Unpaired t-tests were used to assign statistical significance, with ∗p < 0.05 and ∗∗p < 0.01. Data are presented as the mean ± SEM, with individual data points represented by black open circles (glucose, n = 12) or pink open squares (fructose, n = 12). Each circle and square represents an independent biological replicate. In panels E, the data points represent the average of all technical replicates obtained by both operators for a given slide. Panel A was created with BioRender.com/r89w523. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Fig 2: High fructose alters glucose transporter expression, gut morphology, and circulating enterohormones. (A) Mice consumed diets rich in carbohydrates either from glucose or with a portion of glucose replaced by fructose. At the conclusion of the 7-week study protocol (day 52), proximal and distal small intestine sections were collected for gene expression analysis. Gut morphometry was performed blindly by two operators on H&E-stained mid-third jejunum sections. Plasma samples were collected during fasting and following glucose stimulation at week 4, and in a random-fed state at the end of week 7 for enterohormone assessment. (B, C) mRNA expression of sugar transporter genes in proximal and distal small intestine. (D) Representative images of proximal and distal areas of the mid-jejunum. (E, F) Quantification of villus height (v.h.), villus width (v.w.), and crypt depth (c.d.) in proximal and distal gut segments. (G, H) mRNA expression of glucagon (Gng) and glucagon-like peptide (Glp) receptors in proximal and distal small intestine. (I) Fasting and glucose-stimulated Glp2 levels in circulation and (J) area under the Glp2 excursion curve (AUC) at week 4. Circulating levels of (K) Glp2 and (L) Glp1 were also determined in a random-fed state at week 7. Unpaired t-test was used to assign statistical significance, with ∗p < 0.05 and ∗∗∗∗p < 0.0001. Data are presented as mean ± SEM, with individual data points represented by black open circles (glucose, n = 10) or blue open squares (fructose, n = 10). Each circle and square represents an independent biological replicate. In panels E and F, the data points represent the average of all technical replicates obtained by both operators for a given slide. Panel A was created with BioRender.com/u29k943. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
Fig 3: Glp2r inhibition prevents impaired glucose disposal and fatty liver upon high fructose intake. (A) Mice consumed diets rich in carbohydrates, either from glucose or with a portion of glucose replaced by fructose. Mice received daily injections of the glucagon-like peptide (Glp) 2 receptor (Glp2r) antagonist Glp2(3–33) for 23 consecutive days. (B, C) Cumulative food intake and body weight were assessed at weeks 1, 4, and 7. (D, E) Fat and lean mass were determined by nuclear magnetic resonance at week 7. (F) Fasting blood glucose, (G) postprandial glucose tolerance, and (H) the area under the glucose excursion curve (AUC) during OGTT were measured at week 7. (I) Liver triglycerides, and (J) insulin-stimulated total Akt and phosphoAktSer473 levels in the liver, muscle (gastrocnemius), and visceral (gonadal) adipose tissue were assessed in tissues collected at the completion of the 7-week study protocol. Membranes were cut between 55 and 65 kDa, as indicated by dashed or continuous lines. Unpaired t-tests were used to assign statistical significance, with ∗p < 0.05. Data are presented as the mean ± SEM, with individual data points represented by black open circles (glucose, n = 12) or pink open squares (fructose, n = 12). Each circle and square represents an independent biological replicate. Panel A was created with BioRender.com/a80h586. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)
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